Synthesis of New Shogaol Analogues as NRF2 Activators and Evaluation of Their Anti-Inflammatory Activity, Modes of Action and Metabolic Stability.

6-shogaol is a natural and the most potent bioactive vanilloid in dried Zingiber officinale rhizomes. Many scientific studies have reported the diverse biological activities of 6-shogaol. However, the major drawback of 6-shogaol is its instability at room temperature. We synthesised new shogaol thiophene compounds (STCs) by replacing the pentyl group in the sidechain with thiophene derivatives. The STCs were tested for their nuclear factor erythroid 2-related factor 2 (NRF2) activation ability in murine hepatoma cells (Hepa1c1c-7) by determining their NAD(P)H quinone oxidoreductase 1 (NQO1) inducing ability and expression of NRF2-associated antioxidant genes. The anti-inflammatory activity of STCs was determined in Escherichia coli lipopolysaccharide (LPSEc)-stimulated NR2-proficient and -silenced mouse microglial cells (BV-2) by measuring the inflammatory markers, cytokines, and mediators. The modes of action (interacting with the Kelch domain of KEAP1, covalent bonding with cysteines of KEAP1, and inhibition of GSK-3ß enzyme activity) of NRF2 activation by STCs were determined using commercially available kits. The in vitro metabolic stability of the STCs in liver microsomes (humans, rats, and mice) was also investigated. The molecular docking and molecular dynamics studies were conducted to identify the binding poses, stability, and molecular interactions of the STCs in the binding pockets of Kelch and BTB domains of KEAP1 and GSK-3ß enzyme. The new STCs were synthesised in good yields of > 85%, with a purity of about 95%, using a novel synthesis method by employing a reusable proline-proline dipeptide catalyst. The STCs are more potent than 6-shogaol in activating NRF2 and reducing inflammation. The nature of substituents on thiophene has a profound influence on the bioactivity of the STCs. Phenylthiophene STC (STC5) is the most potent, while thiophenes containing electron-withdrawing groups showed weaker bioactivity. The bioactivity of 6-shogaol is in the micromolar range, whereas STC5 showed bioactivity in the sub micromolar range. The STCs showed anti-inflammatory effects via NRF2-dependent and NRF2-independent mechanisms. The STCs improved NRF2 activity through multiple (KEAP1-independent and -dependent) mechanisms. The STCs showed decreased reactivity with thiols than 6-shogaol and thus may possess fewer side-effects than 6-shogaol. The STCs were more metabolically stable than 6-shogaol.

Synthesis and Anti-Inflammatory Activity of 2-Amino-4,5,6,7-tetrahydrobenzo[b]thiophene-Derived NRF2 Activators.

This is the first study investigating the nuclear factor (erythroid-derived 2)-like 2 (NRF2) activity of compounds containing a new scaffold, tetrahydrobenzo[b]thiophene. Eighteen compounds were synthesised and confirmed their NRF2 activation through NQO1 enzymatic activity and mRNA expression of NQO1 and HO-1 in Hepa-1c1c7 cells. The compounds disrupted the interaction between Kelch-like ECH-associated protein 1 (KEAP1) and NRF2 via interfering with the KEAP1's Kelch domain. The compounds exhibited anti-inflammatory activity in Escherichia coli Lipopolysaccharide (LPSEc )-stimulated RAW 264.7 cells. The anti-inflammatory activity of the compounds was associated with their ability to activate NRF2. The compounds reversed the elevated levels of pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α, and IFN-γ) and inflammatory mediators (PGE2, COX-2, and NF-κB). The compounds were metabolically stable in human, rat, and mouse liver microsomes and showed optimum half-life (T1/2 ) and intrinsic clearance (Clint ). The binding mode of the compounds and physicochemical properties were predicted via in silico studies.

Anti-Inflammatory Effects of Auranamide and Patriscabratine-Mechanisms and In Silico Studies.

Auranamide and patriscabratine are amides from Melastoma malabathricum (L.) Smith. Their anti-inflammatory activity and nuclear factor erythroid 2-related factor 2 (NRF2) activation ability were evaluated using Escherichia coli lipopolysaccharide (LPSEc)-stimulated murine macrophages (RAW264.7) and murine hepatoma (Hepa-1c1c7) cells, respectively. The cytotoxicity of the compounds was assessed using a 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. The anti-inflammatory activity was determined by measuring the nitric oxide (NO) production and pro-inflammatory cytokines (Interleukin (IL)-1ß, Interferon (IFN)-γ, tumour necrosis factor (TNF)-α, and IL-6) and mediators (NF-κB and COX-2). NRF2 activation was determined by measuring the nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) quinone oxidoreductase 1 (NQO1), nuclear NRF2 and hemeoxygenase (HO)-1. In vitro metabolic stability was assessed using the mouse, rat, and human liver microsomes. The compounds were non-toxic to the cells at 10 µM. Both compounds showed dose-dependent effects in downregulating NO production and pro-inflammatory cytokines and mediators. The compounds also showed upregulation of NQO1 activity and nuclear NRF2 and HO-1 levels. The compounds were metabolically stable in mouse, rat and human liver microsomes. The possible molecular targets of NRF2 activation by these two compounds were predicted using molecular docking studies and it was found that the compounds might inhibit the Kelch domain of KEAP1 and GSK-3ß activity. The physicochemical and drug-like properties of the test compounds were predicted using Schrodinger small molecule drug discovery suite (v.2022-2).

Durability of SARS-CoV-2 Antibodies from Natural Infection in Children and Adolescents

The authors have withdrawn this manuscript for the following reasons: O_LIOur latest analysis (mid-February, 2022) that includes the addition of 59 children with 3 completed timepoints shows a decrease in N antibodies at T2, versus this preprint that showed an increase in N antibodies from T1 to T2. C_LIO_LIThis preprint did not include those infected with the Omicron variant, which has had a substantial impact on the increase in pediatric infections and has important implications for antibody durability moving forward as such. C_LI Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.

Estimation of Total Immunity to SARS-CoV-2 in Texas

Given the underestimate of seroprevalence in the US due to insufficient testing, accurate estimates of population immunity to SARS-CoV-2 or vaccinations do not exist. Although model-based estimates have been proposed, they require inputting unknown parameters such as viral reproduction number, longevity of immune response, and other dynamic factors. In contrast to a model-based approach for estimating population immunity, or simplistic summing of natural- and vaccine-induced immunity, the current study presents a data-driven statistical procedure for estimating the total immunity rate in a region using prospectively collected serological data along with state-level vaccination data. We present a detailed procedure so that efforts can be replicated regionally to inform policy-making decisions relevant to SARS-CoV-2. Specifically, we conducted a prospective longitudinal statewide cohort serological survey with 10,482 participants and more than 14,000 blood samples beginning on September 30, 2020. Along with Department of State Health Services vaccination data, as of July 4, 2021, the estimated percentage of those with naturally occurring antibodies to SARS-CoV-2 in Texas is 35.3% (95% CI = (33.7%, 36.9%) and total estimated immunity is 69.1%. We conclude the seroprevalence of SARS-CoV-2 is 4 times higher than the state-confirmed COVID-19 cases (8.8%). This methodology is integral to pandemic preparedness.

Swietenine potentiates the antihyperglycemic and antioxidant activity of Metformin in Streptozotocin induced diabetic rats.

Diabetes mellitus or type-2 diabetes, commonly referred as diabetes, is a metabolic disorder that results in high blood sugar level. Despite the availability of several antidiabetic drugs in the market, they still do not adequately regulate blood sugar levels. Thus, in general people prefer to use herbal supplements/medicines along with antidiabetic drugs to control blood sugar levels. One of such herbal medicine is Swietenia macrophylla seeds. It is widely used in Asia for controlling blood sugar levels. One of the major bioactive compounds, Swietenine, is reported to be responsible for controlling blood glucose levels. However, there were no studies on its efficacy in controlling the blood glucose in diabetic rats. In this study, we evaluated the antihyperglycemic activity of Swietenine and its pharmacodynamic interaction with Metformin in Streptozotocin induced diabetes in rats. The activity of Swietenine was investigated at three different doses: 10, 20 and 40 mg/kg body weight (bw). Metformin (50 mg/kg bw) was used as a standard drug. Swietenine (20 and 40 mg/kg bw) and Metformin (50 mg/kg bw) showed significant effect in reducing the glucose, cholesterol, triglycerides, low-density lipoprotein, urea, creatinine, alanine transaminase, alkaline phosphatase, aspartate transaminase, alanine transaminase, and malondialdehyde level in serum while it had increased the high-density lipoprotein, glutathione, and total antioxidant capacity level. In addition, Swietenine (20 and 40 mg/kg) had shown significant synergistic effect with Metformin. Administration of Swietenine at 10 mg/kg bw neither showed activity nor influenced Metformin's activity. The results from this study confirmed the beneficial effects of Swietenine and its synergistic action with Metformin in controlling the dysregulated serum parameters in Streptozotocin induced diabetes in rats.

Carotid endarterectomy for dolichoarteriopathies of internal carotid artery combined with carotid artery severe stenosis / 中国脑血管病杂志

Objective To investigate the clinical efficacy of carotid endarterectomy ( CEA) for the treatment of patients with dolichoarteriopathies of internal carotid artery combined with carotid artery severe stenosis. Methods From January 2013 to February 2018,the clinical data of 18 consecutive patients with dolichoarteriopathy of internal carotid artery combined with carotid artery severe stenosis admitted to the Department of Neurosurgery,the First Affiliated Hospital of Soochow University were analyzed retrospectively. All patients underwent color Doppler ultrasound,CT angiography (CTA),and DSA to assess the diseased vessels before operation,and blood perfusion of the cerebral hemisphere was evaluated by CT perfusion (CTP) imaging. Six patients of Metz grade Ⅱ and 4 of grade Ⅲ underwent valgus CEA +excision of the redundant internal carotid arteries;8 patients of Metz grade I were treated with standard CEA. Postoperative cervical vascular ultrasound,CTA,and CTP examinations were performed in order to understand the vascular patency of the surgery and correction of dolichoarteriopathies of internal carotid artery. The follow-up time was 6 to 72 months. The color Doppler flow imaging and CTA were used to assess the presence or absence of restenosis and MRI was used to evaluate the presence of new cerebral infarction. Results All 18 patients were successfully operated, and they had good vascular patency after operation. The Postoperative CTA showed that the distorted blood vessels had been straightened to varying degrees for ten patients who underwent valgus CEA treatment and the plaques were removed satisfactorily without stenosis for 8 patients treated with standard CEA. After operation,one patient developed sublingual nerve injury symptoms,which was improved after 3 months. One patient developed mild hyperperfusion syndrome,which was improved after 2 weeks. No patients died. Follow-up reexamination showed that all patients had no carotid artery restenosis and new stroke events. Conclusions CEA is a safe and effective treatment for patients with internal carotid artery dolichoarteriopathy combined with severe carotid stenosis. According to the characteristics of the lesions evaluated before surgery, surgical methods should be selected reasonably.

Effect of ischemic preconditioning on expression of intracellular adhesion molecule-1 in brain tissues following traumatic brain injury in rats / 中华创伤杂志

Objective To investigate the effect of ischemic preconditioning (IPC) on expression of intracellular adhesion molecule-1 (ICAM-1) in brain tissues following traumatic brain injury (TBI) in rats.Methods Sixty male SD rats weighing 220-250 g were randomly divided into three groups (n =20 for each):sham operation group,TBI group,and IPC group.Cerebral IPC models were induced by transient occlusion of the bilateral common carotid arteries; TBI models were induced by Feeney's freefalling method; rats in sham operation group were only performed exposure of dura of the right parietal lobe.Ten rats were sacrificed respectively at 6 and 72 hours after TBI and injured brain tissues were harvested to estimate wet/dry weight (W/D) ratio for the brain,determine ICAM-1 expression by immunohistochemistry and perform microscopic examination.Results Brain W/D ratio was significantly increased in TBI group compared with sham operation group (6 h:4.2 ± 0.4 vs 2.7 ± 0.4 ; 72 h:5.0 ±0.1 vs 3.1 ± 0.2,P ＜ 0.05).ICAM-1 expression was up-regulated in TBI group compared with sham operationgroup (6h:25.4±3.5vs8.6±1.3; 72 h:36.5±5.4 vs8.4±1.6,P＜0.05).W/D ratio was significantly decreased in IPC group compared with TBI group (6 h:3.5 ±0.6 vs 4.2 ±0.4; 72 h:3.7 ± 0.4 vs 5.0 ± 0.1,P ＜ 0.05).ICAM-1 expression was down-regulated in IPC group compared with TBI group (6 h:16.5 ± 2.7 vs 25.4 ± 3.5 ; 72 h:24.3 ± 4.6 vs 36.5 ± 5.4,P ＜ 0.05).Milder injury to brain tissues was observed in IPC group than in TBI group.Conclusion IPC can attenuate TBI in rats by down-regulating the expression of ICAM-1.

The Application of Extracranial-Intracranial Vascular Bypass Graft in the Treatment of Intracranial Aneurysms / 国际脑血管病杂志

This article expounds the indication,preoperative ischemic tolerance test,evaluation of collateral circulation,different surgical modes,complications and their prevention in the treatment of intracranial aneurysms during extracranial-intracranial(EC/IC)graft bypass.It is considered that EC/IC bypass graft is an effective therapeutic mode in preventing the occurrence of cerebral ischemia in the treatment of intracranial aneurysms.

Intraoperative microvascular Doppler ultrasonography in urgent clipping surgery for application of ruptured aneurysms / 中国医师进修杂志

Objective To evaluate the efficacy and reliability of intraoperative microvascular Doppler uhrasonography (IMDU)in urgent clipping surgery of ruptured aneurysms,and assess the impact of this method on the surgical procedure itself.Methods For 62 patients who underwent urgent clipping surgery for the treatment of ruptured aneurysms,both blood flow velocities in the aneurismal sac and in the adjacent vessels and alterations of the Doppler spectrum were determined by IMDU before and after aneurysm clipping.The findings of IMDU were analysed and compared with those of visual inspection of the surgical site and postoperative angiography. Results A relevant stenosis of an adjacent vessel induced by efip position that had escaped detection by visual inspection was identified by IMDU in 12 out of 62 patients.In addition, IMDU demonstrated a primarily incomplete clipping aneurysm in 4 out of 62 patients.The clipping aneurysm was repositioned on the basis of the IMDU findings in 16 out of 62 patients.The findings from IMDU equated with those from angiography in 38 eases.Conclusion IMDU is a safe,instantaneous,effective and reliable technique for the urgent dipping surgery of raptured aneurysms.

Decompressive craniectomy and mild hypothermia reduces infarction size and counterregulates Bax and Bcl-2 expression after permanent focal ischemia in rats.

Both mild hypothermia (MH) and decompressive craniectomy (CE) have been shown to have neuroprotective effects in brain ischemia. We investigated a possible effect of MH and a combination of CE and MH (CE + MH) on the changes of infarction size, DNA fragmentation, and immunoreactivities for Bcl-2 and Bax after 24 h of permanent middle cerebral artery occlusion (MCAO) in rats. For the estimation of ischemic brain injury, we calculated the infarct size of the MCA region at 24 h after the MCAO. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin in situ nick labeling (TUNEL) staining was performed for the detection of DNA fragmentation. Immunoreactivities for Bcl-2 and Bax were stained. Infarction size after permanent MCAO was significantly reduced by CE+MH treatment (P < 0.01). Infarction size did not change significantly by application of MH alone (P > 0.05). TUNEL staining was remarkably reduced both in MH-treated animals and in CE + MH-treated animals. Immunoreactivity for Bcl-2 was greatly induced both in MH-treated animals and in CE + MH-treated animals. Induction of immunoreactivity for Bcl-2 was obviously inhibited both in MH-treated animals and in CE + MH-treated animals. It suggests that temporary MH delays infarct evolution and ameliorates neuron apoptosis but does not significantly reduce definite infarction size. CE + MH not only ameliorates neuron apoptosis but also remarkably reduces infarction size.

Arthroscopic reduction and fixation of avulsion fracture of the posterior cruciate ligament from the tibia through an additional posterolateral portal / 中国微创外科杂志

Objective To investigate surgical techniques and results of arthroscopic reduction and fixation for the treatment of avulsion fracture of the posterior cruciate ligament from the tibia through an additional posterolateral portal. Methods An additional posterolateral portal was established by using the inside-out technique under arthroscopic view.After a protective sleeve was placed through the portal and the displaced fracture was reduced,the fragment was fixed by the guide wire drilled through the sleeve temporarily.If the fluoroscopic control showed an anatomic reduction of the fragment and good placement of the wire,the self-attacked cannulated screw and washer was placed intra-articularly over the guide wire through the sleeve for the directive internal fixation of the avulsion fracture.Aggressive rehabilitation programs were recommended postoperatively.Results Except 1 case with 11 mm displacement and rotation of the fragment treated by the arthrotomy after the failure of arthroscopic reduction,the arthroscopic operation was accomplished in all the remaining 10 cases.The operative time was 63～98 min(mean,87.3 min).No injuries of popliteal vessels or nerves occurred.The postoperative X-ray firms showed an anatomic reduction of the fragment and good placement of the screw and washer.With the negative posterior drawer test confirmed by the physical examination,normal range of motion of the injured knee joint and gait were achieved in the 10 patients at 4～7 weeks after surgery.The bone union was confirmed by X-ray films at the 3 months postoperatively.The computer KT-2000 arthrometer measurements of posterior tibial displacement of both knee joints showed side-to-side difference not more than 1.2 mm in 6 patients at 5 months after surgery.Conclusions The additional posterolateral portal can be established by the arthroscopic inside-out technique safely.The displaced fragment of the avulsion fracture of the posterior cruciate ligament from the tibia can be reduced and fixed with the self-attacked cannulated screw and washer arthroscopicly.

The role of nitric oxide and nitric oxide synthase in killing tumor cells / 第二军医大学学报

Objective: To study the role of inducible nitric oxide synthase (iNOS) in activated macrophages and nitric oxide (NO) in the defence against tumors. Methods: Macrophages were obtained by alveolar lavages of mice and activated through incubation with recombinant murine INF? in vitro. P815 cells were added to the culture. Culture supernates were collected to measure the activity of iNOS and NO. Tumoricidal activity of macrophages was determined in presence and absence of the specific inhibitor of NO synthase: L NMMA. Results: NO production and activity of iNOS induced by activated macrophages were positively related with concentration of INF? in macrophage P815 coculture. The addition of L NMMA to the culture suppressed NO production, the inhibitory rate of activated macrophages against P815 cells was reduced distinctly ( P

THE RECEPTOR ROLE OF GLYCOPHORIIN A DURING THE INVASION OF HUMAN ERYTHROCYTE BY PLASMODIUM FALCIPARUM / 中国寄生虫学与寄生虫病杂志

The effects of GPA, anti-GPA IgG, ?1acid glycoprotcin (?1-AGP), ovomucoid (OM) and wheat germ agglutinin (WGA) on the invasion of erythrocytes by P. falciparum were obseryed in vitro. GPA, anti-GPA IgG and WGA at low concentrations had obviously inhibitory effects on thc invasion. There were hyperbolic relationships between the concentrations of these inhibitors and their inhibition rates. OM, ?1-AGP and non-spccific anti-serum had no obvious effect on the invasion. It is confirmed for the first time from the biological characteristics of receptor-ligand interaction that thc combination of GPA with P. falciparum merozoites has high spccificity, high affinity with a tendency to sa-turation and can produce specific biological effects.